Use of metalloprotinease inhibitors to inducing and/or stimulating growth of hair or hair or hairs and/or for slowing down their loss

ABSTRACT

The invention concerns the use in/for the preparation of a composition, as active principle in a physiologically acceptable medium, an efficient amount of at least a metalloproteinase inhibitor or any functional biological equivalent, for inducing and/or stimulating growth of hair or hairs and for slowing down their loss. The invention also concerns a method for the cosmetic treatment of hair or hairs.

The present invention relates to the use, in or for the preparation of acomposition, as active principle in a physiologically acceptable medium,of an effective amount of at least one metalloprotease inhibitor, or ofany functional biological equivalent, which is intended to induce and/orstimulate the growth of head hair or other hairs and/or to slow downtheir loss.

In humans, the growth and renewal of the hair are mainly determined bythe activity of the hair follicles and by their dermo-epidermalenvironment. Their activity is cyclic and essentially comprises threephases, i.e. the anagenic phase, the catagenic phase and the telogenicphase.

The active anagenic phase or growth phase, which lasts for several yearsand during which the hair gets longer, is followed by a very short andtransient catagenic phase which lasts a few weeks, and then comes a restphase, known as the telogenic phase, which lasts a few months.

At the end of the rest period, the hair falls out and another cyclebegins. The head of hair is thus under constant renewal, and out of theapproximately 150,000 hairs which make up a head of hair, at any givenmoment, approximately 10% of them are at rest and will thus be replacedwithin a few months.

In a large number of cases, early hair loss occurs in individuals whoare genetically predisposed, and it usually affects men. This moreparticularly concerns androgenetic or androgenic or evenandrogenogenetic alopecia.

This alopecia is essentially due to a disruption in hair renewal whichleads, in a first stage, to an acceleration of the frequency of thecycles, at the expense of the quality of the hair and then at theexpense of its quantity. A gradual depletion of the head of hair takesplace by regression of the so-called “terminal” hairs at the downystage. Regions are preferentially affected, in particular the temples orfrontal bulbs and the back of the head in men, whilst in women diffusealopecia of the vertex is observed.

Substances for suppressing or reducing alopecia, and in particular forinducing or stimulating hair growth or reducing hair loss, have beensought for many years in the cosmetics and pharmaceutical industries.

Admittedly, in this respect, a large number of very diverse activecompounds have already been proposed, such as, for example,2,4-diamino-6-piperidinopyrimidine 3-oxide or “Minoxidil” described inU.S Pat. No. 4,596,812, or the many derivatives thereof, such as thosedescribed, for example, in patent applications EP 0 353 123, EP 0 356271, EP 0 408 442, EP 0 522 964, EP 0 420 707, EP 0 459 890 and EP 0 519819.

Mention may also be made of 6-amino-1,2-dihydro1hydroxy-2-imino-4-piperidinopyrimidine and its derivatives, which aredescribed more particularly in patent U.S Pat. No. 4,139,619.

However, it would generally still be advantageous and useful to be ableto provide active compounds other than those already known.

Now, after considerable research conducted in this matter, the Applicanthas just discovered that a metalloprotease inhibitor, or any functionalbiological equivalent, makes it possible to induce and/or stimulate thegrowth of head hair or other hairs, and/or to reduce their loss in aneffective manner.

Metalloproteases (MMPs) are members of a family of proteolitic enzymes(endoproteases) which contain a zinc atom coordinated to 3 cysteineresidues and one methionine residue in their active site and whichdegrade the macromolecular components of the extracellular matrix andthe basal sheets at neutral pH (collagen, elastin, etc.). These enzymes,which are very widely distributed in the living world, are present, butweakly expressed, in normal physiological situations such as organgrowth and tissue renewal. However, their overexpression in man andtheir activation are associated with many processes which involve thedestruction and remodelling of the matrix. This entails, for example, anuncontrolled resorption of the extracellular matrix.

Metalloproteases are produced and secreted in an inactive zymogenic form(pro-enzyme). These zymogenic forms are then activated in theextracellular environment by the removal of a propeptide region. Themembers of this family can activate each other.

Regulation of the activity of MMPs thus takes place at the level of theexpression of the genes (transcription and translation), at the level ofthe activation of the zymogenic form, or at the level of the localcontrol of the active forms.

The main regulators of the activity of MMPs are the tissue inhibitors ofmetalloproteases, or TIMPs. However, the expression of MMPs is alsomodulated by growth factors, cytokines, oncogenic products (ras, jun) ormatrix constituents.

The metalloprotease family consists of several well-defined groups basedon their resemblances in terms of substrate specificity and structure(see Woessner J. F., Faseb Journal, vol. 5, 1991, 2145). Among thesegroups, mention may be made of collagenases intended to degradefibrillar collagens (MMP-1 or interstitial collagenase, MMP-8 orneutrophil collagenase, and MMP-13 or collagenase 3), gelatinases whichdegrade type IV collagen or any form of denatured collagen (MMP-2 orgelatinase A (72 kDa), MMP-9 or gelatinase B (92 kDa)), stromelysinswhose broad spectrum of activity applies to extracellular matrixproteins such as glycoproteins (fibronectin, laminin), proteoglycans,etc. or alternatively membrane metalloproteases.

The Applicant has now discovered that metalloproteases are present inthe internal structures of hair follicles, namely in the innerepithelial sheath (IRS). In particular, MMP-9 is found in the IRS.

Now, it is known that in the course of the hair cycle, hair folliclespass from a low-level location in the dermis in the anagenic phase, to ahigh-level location in the dermis during the telogenic phase. Thismovement should be accompanied by a change in the extracellular matrixwhich allows the migration of the follicle, this change possibly beingdue to an expression of the MMPs, bringing about a controlleddegradation of the said extracellular matrix. It is at the end of thetelogenic phase that hair loss occurs. However, it is also known thatcytokines and growth factors have an influence on the hair cycle. Forexample, epidermal growth factor (EGF) promotes the in vitro transitionfrom the anagenic phase to the catagenic phase (formation of a “club”structure characteristic of the catagenic phase), this being the phasewhich precedes the loss of the head hairs or other hairs. It is alsoknown, as the Applicant has demonstrated, that there is an inflammatoryphase in alopecia.

The Applicant has shown that the MMPs and particularly MMP-9, can beinduced by interleukin-1 and/or EGF, in particular in the fibroblasts ofthe dermal papillae.

The advantage of reducing the expression of MMPs in the scalp in orderto slow down or inhibit the degradation of the perifollicular matrix andthus to slow down or even prevent hair loss may thus be appreciated.

The Applicant thus proposes the use of metalloprotease inhibitors toinduce and/or stimulate the growth of head hair or other hairs and/or toslow down their loss.

Thus, the invention relates to the use, in or for the preparation of acomposition, of an effective amount of at least one metalloproteaseinhibitor or of any functional biological equivalent, which is intendedto induce and/or stimulate the growth of head hair or other hairs and/orto slow down their loss.

The expression “functional biological equivalent” means any moleculewhich is functionally equivalent in terms of biological function, atleast one of the components of which may have been changed for anequivalent component.

Examples which may be mentioned are peptides, one biological equivalentof which may be a peptide in which at least one amino acid residue hasbeen replaced with another amino acid having a similar hydropathicindex.

The expression “metalloprotease inhibitor” means any molecule capable ofregulating the activity of MMPs either at the level of the expression ofthe genes (transcription and translation) or at the level of theactivation of the zymogenic form of MMPS, or alternatively at the levelof the local control of the active forms.

The main regulators of the activity of MMPs are natural moleculespresent in the tissues, known as tissue inhibitors of metalloproteases,or TIMPs.

However, these inhibitors may also be known chemical molecules such as,for example, hydroxamic acid derivatives, cation-chelating agents,growth factors, cytokines, oncogenic products (ras, jun) or matrixconstituents.

According to the invention, tissue inhibitors of metalloproteases(TIMPs) such as, for example, the peptides known in the prior art underthe names TIMP-1, TIMP-2, TIMP-3 and TIMP-4 (Woessner J. F., FasebJournal, 1991) are preferably used.

Thus, the invention relates more particularly to the use, in or for thepreparation of a composition, of an effective amount of at least onetissue inhibitor of metalloproteases (TIMP) or of any functionalbiological equivalent, the inhibitor or the composition eing intended toinduce and/or stimulate the growth of head hair or other hairs and/or toslow down their loss.

The peptides known in the prior art under the names TIMP-1, TIMP-2,TIMP-3 and TIMP-4 are preferably used according to the invention astissue inhibitors of metalloproteases.

Needless to say, according to the invention, the metalloproteaseinhibitors can be used alone or as a mixture.

It may be the case that, for reasons of resistance to degradation, it isnecessary according to the invention to use a protected form of themetalloprotease inhibitor. The form of the protection should obviouslybe a biologically compatible form. Many biologically compatible forms ofprotection may be envisaged, such as, for example, acylation oracetylation of the amino-terminal end or amidation of thecarboxy-terminal end.

Thus, the invention relates to a use as defined above, characterized inthat the metalloprotease inhibitor is in a protected or unprotectedform.

A protection based either on the acylation or acetylation of theamino-terminal end, or on the amidation of the carboxy-terminal end, oralternatively on both approaches, is preferably used according to theinvention.

Among the chemical inhibitors which may be mentioned are thiols andhydroxamates.

The amount of metalloprotease inhibitor which can be used according tothe invention obviously depends on the desired effect and should be inan amount which is effective to induce and/or stimulate the growth ofhead hair or other hairs and/or to slow down their loss.

By way of example, the amount of metalloprotease inhibitor which can beused according to the invention may range, for example from 0.01% to 5%and preferably from 0.05% to 2% relative to the total weight of thecomposition.

The composition is preferably a cosmetic composition.

The composition according to the invention can be administered enterallyor parenterally. preferably, via the parenteral route, the compositionis administered topically.

The physiologically acceptable medium in which the peptide is usedaccording to the invention may be anhydrous or aqueous. The expression“anhydrous medium” means a solvent medium containing less than 1% water.This medium may consist of a solvent or a mixture of solvents chosenmore particularly from C₂-C₄ lower alcohols such as ethyl alcohol,alkylene glycols such as propylene glycol, and alkylene glycol alkylethers or dialkylene glycol alkyl ethers, the alkyl or alkylene radicalsof which contain from 1 to 4 carbon atoms. The expression “aqueousmedium” means a medium consisting of water or of a mixture of water andanother physiologically acceptable solvent, chosen in particular fromthe organic solvents mentioned above. In this last case, when theseother solvents are present, they represent approximately 5% to 95% byweight of the composition.

It is possible for the physiologically acceptable medium to containother adjuvants usually used in cosmetics, such as surfactants,thickeners or gelling agents, cosmetic agents, preserving agents, andacidifying and basifying agents that are well known in the prior art,and in amounts that are sufficient to obtain the desired presentationform, in particular a more or less thickened lotion, a gel, an emulsionor a cream. The composition can optionally be used in a form pressurizedas an aerosol or vaporized from a pump-dispenser bottle.

It is also possible for the peptide to be used in combination withcompounds for further improving the activity on hair regrowth and/or onslowing down hair loss, which have already been described for thisactivity.

Among the latter compounds, mention may be made more particularly, in anon-limiting manner, of:

nicotinic acid esters, in particular including tocopheryl nicotinate,benzyl nicotinate and C₁-C₆ alkyl nicotinates such as methyl or hexylnicotinate;

pyrimidine derivatives, such as6-amino-1,2-dihydro-1-hydroxy-2-imino-4-piperidinopyrimidine also knownas “Minoxidil” and as described in U.S. Pat. No. 4 139 619;

antiandrogenic agents

5-reductase inhibitors;

OH-radical scavengers, such as dimethylsulfoxide;

peptides such as, for example, the tripeptide Lys-Pro-Val;

microorganism extracts, particularly bacterial extracts;

plant extracts.

Other compounds can also be added to the above list, namely, forexample, phospholipids such as lecithin, linoleic acid, linolenic acid,salicylic acid and derivatives thereof described in French patent FR 2581 542, for instance salicylic acid derivatives bearing an alkylradical containing from 2 to 12 carbon atoms in position 5 of thebenzene ring, hydroxycarboxylic or ketocarboxylic acids and estersthereof, lactones and the corresponding salts thereof, carotenoids,eicosatetraenoic and eicosatrienoic acids or esters and amides thereof,vitamin D and derivatives thereof.

The cosmetic composition according to the invention can be applied tothe alopecic regions of the scalp and hair of an individual, and isoptionally left in contact for several hours and is optionally to berinsed out. For example, the cosmetic composition containing aneffective amount of at least one metalloprotease inhibitor can beapplied to the hair and the scalp in the evening, kept in contactthroughout the night and optionally shampooed out in the morning. Theseapplications can be repeated daily for one or more months depending onthe individual.

Thus, a subject of the present invention is also a cosmetic process fortreating the hair and/or the scalp, characterized in that it consists inapplying a cosmetic composition containing an effective amount of atleast one metalloprotease inhibitor to the hair and/or the scalp, inleaving this composition in contact with the hair and/or the scalp, andoptionally in rinsing it off.

The treatment process has the characteristics of a cosmetic processsince it improves the aesthetics of the hair by making it more vigorousand making it look better.

Examples will now be given by way of illustration, which should not inany way limit the scope of the invention.

EXAMPLE 1 Daily Lotion

TIMP-1 0.01 g TIMP-2 0.01 g 2,4 Diaminopyrimidine 3-oxide 0.75 g 95°ethanol 30 g Fragrance qs Dyes qs Demineralized water qs 100 g

EXAMPLE 2 Liposomal Gel

Natipide II^({circle around (1)}) (i.e. 2 g of 10 g phospholipids)TIMP-2 0.025 g Carbomer 0.25 g Triethanolamine qs pH = 7 Preservingagents qs Demineralized water qs 100 g^({circle around (1)})Water/Alcohol/Lecithin mixture from the companyNattermann

What is claimed is:
 1. A method for inducing and/or stimulating thegrowth of hair and/or slowing hair loss comprising applying to the hairand/or the scalp of an individual in need thereof, a topical compositioncomprising an effective amount of at least one tissue inhibitor ofmetalloproteases (TIMP) or a protected form thereof in a physiologicallyacceptable medium, leaving said composition in contact with the hairand/or the scalp for a sufficient time and optionally rinsing saidcomposition from the treated area.
 2. The method according to claim 1,wherein said at least one tissue inhibitor of metalloproteases is ofnatural origin.
 3. The method according to claim 2, wherein said atleast one tissue inhibitor of metalloproteases is a protected formthereof.
 4. The method according to claim 3, wherein said protected formis protected by acylation or acetylation of the amino-terminal end. 5.The method according to claim 3, wherein said protected form isprotected by arnidation of the carboxy-terminal end.
 6. The methodaccording to claim 3, wherein said protected form is protected byacylation or acetylation of the amino-terminal end and by amidation ofthe carboxy-terminal end.
 7. The method according to claim 1, whereinsaid at least one tissue inhibitor of metalloproteases is present in anamount of between 0.01% and 5% relative to the total weight of thecomposition.
 8. The method according to claim 1, wherein said at leastone tissue inhibitor of metalloproteases is present in an amount ofbetween 0.05% and 2% relative to the total weight of the composition. 9.The method according to claim 1, wherein said at least one tissueinhibitor of metalloproteases is of synthetic origin.
 10. A method forinducing and/or stimulating the growth of hair and/or slowing hair losscomprising applying to the hair and/or the scalp of an individual inneed thereof, a topical composition comprising an effective amount of atleast one tissue inhibitor of metalloproteases (TIMP) selected from thegroup consisting of TIMP-1, TIMP-2, TIMP-3, TIMP-4 and the protectedforms thereof in a physiologically acceptable medium, leaving saidcomposition in contact with the hair and/or the scalp for a sufficienttime and optionally rinsing said composition from the treated area. 11.The method according to claim 10, wherein said composition comprisesTIMP-1.
 12. The method according to claim 11, wherein said TIMP-1 ispresent in an amount of between 0.01% and 5 % relative to the totalweight of the composition.
 13. The method according to claim 10, whereinsaid composition comprises TIMP-2.
 14. The method according to claim 13,wherein said TIMP-2 is present in an amount of between 0.01% and 5%relative to the total weight of the composition.
 15. The methodaccording to claim 10, wherein said composition comprises a mixture oftissue inhibitors of metalloproteases.
 16. The method according to claim15, wherein said mixture of tissue inhibitors of metalloproteases ispresent in an mount of between 0.01% and 5% relative to the total weightof the composition.
 17. The method according to claim 15, wherein saidcomposition comprises a mixture of TIMP-1 and TIMP-2.
 18. The methodaccording to claim 17, wherein said mixture of TIMP-1 and TIMP-2 ispresent in an amount of between 0.01% and 5% relative to the totalweight of the composition.
 19. The method according to claim 10, whereinsaid at least one tissue inhibitor of metalloproteases is a protectedform thereof.
 20. The method according to claim 19, wherein saidprotected form is protected by acylation or acetylation of theamino-terminal end.
 21. The method according to claim 20, wherein saidprotected form is protected by amidation of the carboxy-terminal end.22. The method according to claim 21, wherein said protected form isprotected by acylation or acetylation of the amino-terminal end and byamidation of the carboxy-terminal end.
 23. The method according to claim10, wherein said at least one tissue inhibitor of metalloproteases ispresent in an amount of between 0:01% and 5% relative to the totalweight of the composition.
 24. The method according to claim 10, whereinsaid at least one tissue inhibitor of metalloproteases is present in anamount of between 0.05% and 2% relative to the total weight of thecomposition.